摘要
以携带pCAMBIA1301(含GUS基因)质粒的EHA105菌株侵染玉米优良自交系齐319幼胚诱导的胚性愈伤组织,共培养5 d(特殊试验除外)后,对GUS基因瞬时表达率影响玉米转化的共培养环境进行研究。结果表明,玉米转化效率随共培养基pH降低而升高,临界共培养pH为5.2;共培养基中添加200μmol/L乙酰丁香酮(AS)对提高玉米愈伤组织的转化率是必要的;共培养3和5 d的GUS表达率呈现先升高后降低的趋势,以25℃共培养3 d的GUS表达率最高,为33.74%。
An optimized transformation system was developed for maize elite inbred lines using Agrobacterium tumefaciens mediated gene transfer by studying various important co-culture systems that affect the efficiency of genetic transformation.The system was performed by transient expression of GUS gene after embryogenic callus co-cultured for five days（except special experiment）.The study using the embryogenic callus derived from immature embryos of maize inbred line Qi 319 showed that the genetic transformation efficiency was increased with the decreasing of co-cultivation medium pH.The appropriate pH of co-cultivation medium was 5.2.It was beneficial to add acetosyringone（AS） into co-cultivation medium for improving transformation efficiency.The result also indicated that the transformation efficiency after adding 200 μmol/L acetosyringone（AS） into the co-cultivation medium was significantly higher than other treatments.Both of the GUS gene expression efficiencies co-cultured for three days and five days were increased at first and then decreased.The highest expression efficiency of GUS gene could reach 33.74% when co-cultured three days in dark at 25 ℃.
出处
《西北农业学报》
CAS
CSCD
北大核心
2011年第9期40-42,共3页
Acta Agriculturae Boreali-occidentalis Sinica
基金
辽宁省玉米育种工程技术分中心项目(2006402031)
沈阳市特种玉米遗传生理重点实验室项目(1062107-1-00)
关键词
玉米
共培养
农杆菌介导
遗传转化
Maize
Co-culture systems
Agrobacterium tumefaciens mediated
Genetic transformation
